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Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM)
Author(s) -
Hanley Quentin S.,
Lidke Keith A.,
Heintzmann Rainer,
ArndtJovin Donna J.,
Jovin Thomas M.
Publication year - 2005
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20177
Subject(s) - optical sectioning , fluorophore , microscope , förster resonance energy transfer , fluorescence lifetime imaging microscopy , microscopy , fluorescence , confocal , optics , confocal microscopy , materials science , light sheet fluorescence microscopy , fluorescence microscope , focus (optics) , physics
Background The programmable array microscopes (PAMs) are a family of instruments incorporating arbitrary control of the patterns of illumination and/or detection. The PAM can be used in sectioning and nonsectioning modes, thereby constituting a useful platform for fluorescence lifetime imaging. Methods and Results We used a PAM for acquisition of optically sectioned and widefield fluorescence lifetime images, in which contrast was increased predominantly by suppressing out‐of‐focus light contributions. We simulate, display, and discuss the effects of blurring and fluorophore heterogeneity on lifetime imaging in widefield and confocal configurations. Conclusion Sectioning improves the quality of lifetime images of samples with multiple fluorophores or spatially varying Förster resonance energy transfer. © 2005 International Society for Analytical Cytology.